• Toxicological Research
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• 제30권2호
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• pp.121-130
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• 2014

The larval form of Tenebrio molitor (T. molitor) has been eaten in many countries and provides benefits as a new food source of protein for humans. However, no information exists regarding its safety for humans. The objective of the present study was to evaluate the genotoxicity and repeated dose oral toxicity of the freeze-dried powder of T. molitor larvae. The genotoxic potential was evaluated by a standard battery testing: bacterial reverse mutation test, in vitro chromosome aberration test, and in vivo micronucleus test. To assess the repeated dose toxicity, the powder was administered once daily by oral gavage to Sprague-Dawley (SD) rats at dose levels of 0, 300, 1000 and 3000 mg/kg/day for 28 days. The parameters which were applied to the study were mortality, clinical signs, body and organ weights, food consumption, ophthalmology, urinalysis, hematology, serum chemistry, gross findings and histopathologic examination. The freezedried powder of T. molitor larvae was not mutagenic or clastogenic based on results of in vitro and in vivo genotoxicity assays. Furthermore, no treatment-related changes or findings were observed in any parameters in rats after 28 days oral administration. In conclusion, the freeze-dried powder of T. molitor larvae was considered to be non-genotoxic and the NOAEL (No Observed Adverse Effect Level) was determined to be 3000 mg/kg/day in both sexes of SD rats under our experimental conditions.

• 한국축산식품학회지
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• 제37권3호
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• pp.351-359
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• 2017

This study was conducted to investigate the physicochemical properties of meat batters prepared with fresh pork meat, back fat, water, and salt and formulated with three different amounts (5%, 10%, and 15%) of silkworm pupae (Bombyx mori) powder and transglutaminase (TG). Meat batters formulated with silkworm pupae powder showed significantly higher contents of protein and ash than control batter. Addition of silkworm pupae to batter also showed significantly lower cooking loss than the control. Moreover, meat batter containing 15% silkworm pupae showed no significant difference in redness value compared to the control. In addition, pH, viscosity, hardness, gumminess, and chewiness were improved after the addition of silkworm pupae. Furthermore, meat batter formulated with TG and silkworm pupae showed improved hardness, gumminess, chewiness and viscosity compared to control batter. Addition of 1% TG with 15% silkworm pupae to meat batter resulted in significantly higher pH, textures, and viscosity. Our data suggest that both silkworm pupae and TG can be added to meat batter to improve its physicochemical properties. Therefore, combination of silkworm pupae and TG could be a new nutritional and functional source for meat products.

• Asian-Australasian Journal of Animal Sciences
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• 제23권12호
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• pp.1587-1593
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• 2010

Pastures of the Apennines of Central Italy contribute to feed resources of high altitude (above 1,300 m sea level) grazing systems. The objective of this study was to evaluate the effectiveness of faecal extract from the yak (Bos grunnienes) as an alternative microbial inoculum to rumen fluid for estimation of digestibility of several forage species. Forage samples produced at high altitude were tested in this study: four legumes (Lathyrus sativus L., Lotus corniculatus L., Onobrychis viciaefolia L. and Trifolium pratense L.), three forbs (Achillea millefolium L., Potentilla reptans L. and Teucrium flavum L.) and one grass (Brachipodyum pinnatum L.) were incubated with yak rumen fluid or faecal extract. A large variability in chemical composition was observed among the species collected. Rumen liquor and faecal samples were collected from adult healthy yak. The $Daisy^{II}$ incubator was used to evaluate the nutrient digestibility of forages using rumen liquor as control and faecal extract as alternative microbial inoculum sources. Filter bags containing samples of browse species were added to the four digestion vessels along with their respective inoculum and then incubated for 48 h and dry matter (DM), organic matter (OM), crude protein (CP), neutral and detergent fiber (NDF) digestibility was determined. There was a significant relationship between estimates, indicating that faecal liquor has the potential to be used instead of rumen fluid for estimation of in vitro digestibility of plants. It is concluded that the $Daisy^{II}$ incubator results are appropriate for the determination of in vitro digestibility of nutrients using faecal liquor to define the potential for adaptation of yak to new pastures.

• Bulletin of the Korean Chemical Society
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• 제23권8호
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• pp.1139-1143
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• 2002

Analysis of lysophosphatidic acids (LPAs) is of clinical importance as they can serve a potential marker for ovarian and other gynecological cancers and obesity. It is critically important to develop a highly sensitive and specific method for the early detection of gynecological cancers to improve the overall outcome of this disease. We have established a novel quantification method of LPAs in human plasma by negative ionization tandem mass spectrometry (MS-MS) using multiple reaction monitoring (MRM) mode without the conventional TLC step. Protein-bound lipids, LPAs in plasma were extracted with methanol : chloroform (2:1) containing LPA C14:0 as an internal standard under acidic condition. Following back extraction with chloroform and water, the centrifuged lower phase was evaporated and reconstituted in methanol. The reconstituted solution was directly injected into electrospray source of MS/MS. For MRM mode, Q1 ions selected were m/z 409, 433, 435, 437 and 457 which corresponds to molecular mass [M-H]- of C16:0, C18:2, C18:1, C18:0 and C20:4 LPA, respectively. Q2 ions selected for MRM were m/z 79, phosphoryl product. Using MS/MS with MRM mode, all the species of LPAs were completely separated from plasma matrix without severe interferences. This method allowed simultaneous detection and quantification of different species of LPAs in a plasma over a linear dynamic range of 0.01-25 ㎛olL-1 . The detection limit of the method was 0.3 pmol/mL, with a correlation coefficient of 0.9983 in most LPAs analyzed. When applied to the plasmas of normal and gynecological cancer patients, this new method differentiated two different groups by way of total LPA level.

• BMB Reports
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• 제40권1호
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• pp.15-21
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• 2007

Breast cancer is the most common malignancy among women, and mutations in the BRCA1 gene produce increased susceptibility to these malignancies in certain families. In this study, the forward 1-13 exons of breast cancer associated gene BRCA1 were cloned from breast cancer cell line ZR-75-30 by RT-PCR method. Sequence analysis showed that nine BRCA1 splice forms were isolated and characterized, compared with wild-type BRCA1 gene, five splice forms of which were novel. These splice isoforms were produced from the molecular mechanism of 5' and 3' alternative splicing. All these splice forms deleting exon 11b and the locations of alternative splicing were focused on two parts:one was exons 2 and 3, and the other was exons 9 and 10. These splice forms accorded with GT-AG rule. Most these BRCA1 splice variants still kept the original reading frame. Western blot analysis indicated that some BRCA1 splice variants were expressed in ZR-75-30 cell line at the protein level. In addition, we confirmed the presence of these new transcripts of BRCA1 gene in MDA-MB-435S, K562, Hela, HLA, HIC, H9, Jurkat and human fetus samples by RT-PCR analysis. These results suggested that breast cancer associated gene BRCA1 may have unexpectedly a large number of splice variants. We hypothesized that alternative splicing of BRCA1 possibly plays a major role in the tumorigenesis of breast and/or ovarian cancer. Thus, the identification of cancer-specific splice forms will provide a novel source for the discovery of diagnostic or prognostic biomarkers and tumor antigens suitable as targets for therapeutic intervention.

• Plant Breeding and Biotechnology
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• 제6권4호
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• pp.413-425
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• 2018

A previous work developed and identified a new omega-5 gliadin deficient wheat line named O-free by crossing Keumkang and Olgeuru, which is nutritionally quite meaningful in that omega-5 gliadin is one of the known wheat allergens. To verify the characteristics of the O-free, we performed RNA sequencing (RNAseq) analysis of the O-free and the two parent lines (Keumkang and Olgeuru). The results of the similarity analysis with the ESTs for gliadins and glutenins showed that the O-free ESTs had no similarity with the omega-5 gliadin sequences but had similarity to other gliadins and glutenins. Furthermore, mapping results between the raw RNAseq data from the O-free and the omega-5 gliadin sequence showed a clear deletion of the N-terminal sequences which are an important signature of omega-5 gliadin. We also designed specific PCR primers that could identify omega-5 gliadin in the genomic DNA. The results showed that no omega-5 gliadin fragments were detected in the O-free. According to these results, we confirmed that the deficiency of omega-5 gliadin in the O-free is not caused by post-transcriptional or post-translational regulations such as epigenetic phenomena but by a simple deletion in the chromosome. Furthermore, we showed that the low-molecular weight glutenin subunit (LMW-GS) gene in the O-free had a single nucleotide polymorphism (SNP) causing a premature stop codon, resulting in a truncated polypeptide. We expect that the O-free line may serve as an excellent source of wheat that could prevail in the hypo-allergen wheat market, which has recently gained interest world-wide.

• International Journal of Industrial Entomology and Biomaterials
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• 제45권2호
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• pp.84-92
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• 2022

White-Jade silkworm (previously also known as Baegokjam) variety is the most popular silkworm variety that produces white cocoons. In 2021, the market share of White-Jade variety in Korea is very high, accounting for 88% of the silkworm production. Daebaekjam, Dodamsilkworm, Kumkangjam, and Kumokjam varieties, which have recently been established, make white cocoons like White-Jade. In this study, we found that 5 types of HongJams produced from 5 varieties of silkworms producing white cocoons did not show any severe difference in proximate analysis. The amounts of crude proteins, the most abundant nutrient, were between 71.05 ~ 73.38%, and those of crude lipids were 13.89 ~ 14.69% in 5 types of HongJams. In addition, there was no difference between White-Jade HongJam (WJ) and Daebaekjam HongJam in amino acid compositions. The amount of unsaturated fatty acid was significantly higher in WJ than in the other four types of HongJams, but the omega-6 fatty acids/omega-3 fatty acids ratio was higher in the four types of HongJams. Most of the minerals were higher in four types of HongJams than in WJ, and three heavy metal were not detected in all 5 types of HongJams. Phytochemicals were also most abundant in WJ, but the difference in the amounts were not severe. And pepsin digestibility was the highest for Kumokjam HongJam and the lowest for Dodam-silkworm HongJam, but the difference was not severe. The nutritional component analysis results of this study suggested that four new varieties can be used for producing HongJams, and Daebaekjam can replace White-Jade the most as the protein source.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2001년도 추계학술대회
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• pp.39-45
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• 2001

Extremophiles are unique microorganisms that are adapted to survive in ecological niches such as high or low temperatures, extremes of pH, high salt concentrations and high pressure. These unusual microorganisms have unique biochemical features which can be exploited for use in the biotechnological industries. Due to the high biodiversity of extremophilic archaea and bacteria and their existence in various biotopes a variety of biocatalysts with different physicochemical properties have been discovered. The extreme molecular stability of their enzymes, membranes and the synthesis of unique organic compounds and polymers make extremophiles interesting candidates for basic and applied research. Some of the enzymes from extremophiles, especially hyperthermophilic marine microorganisms (growth above $85^{\circ}C$), have already been purified in our laboratory. These include the enzyme systems from Pyrococcus, Pyrodictium, Thermococcus and Thermotoga sp. that are involved in polysacharide modification and protein bioconversion. Only recently, the genome of the thermoalkaliphilic strain. Anaerobranca gottschalkii has been completely sequenced providing a unique resource of novel biocatalysts that are active at high temperature and pH. The gene encoding the branching enzyme from this organism was cloned and expressed in a mesophilic host and finally characterized. A novel glucoamylase was purified from an aerobic archaeon which shows optimal activity at $90^{\circ}C$ and pH 2.0. This thermoacidophilic archaeon Picrophilus oshimae grows optimally at pH 0.7 and $60^{\circ}C$. Furthermore, we were able to detect thermoactive proteases from two anaerobic isolates which are able to hydrolyze feather keratin completely at $80^{\circ}C$ forming amino acids and peptides. In addition, new marine psychrophilic isolates will be presented that are able to secrete enzymes such as lipases, proteases and amylases possessing high activity below the freezing point of water.

• Journal of Animal Science and Technology
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• 제66권2호
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• pp.398-411
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• 2024

Upregulation of the nutritional value of feed is the major target of various studies in the livestock industry, and dietary enzyme supplementation could aid in digesting the nondegrading nutrients of grains in feed ingredients. Dried distillers' grains with solubles (DDGS) is a byproduct of the fermentation process in the beverage industry and can be used as a large supply source of fiber in feed. Therefore, we conducted an experiment with male broiler chickens to investigate the effect of various types of enzymes on DDGS and compare the efficacy of single enzyme and multienzyme complexes on growth performance and gut environments in broiler chickens. We used 420 1-day-old broiler chickens (Ross 308), and they were allotted into 4 dietary treatments with seven replications (CON, corn-soybean meal [SBM] diet; NC, DDGS supplemented diet; SE, 0.05 % of mannanase supplemented DDGS-based diet; MC, 0.10% of multienzyme complex (mannanase and xylanase, glucanase) supplemented DDGS-based diet. The dietary exogenous enzyme in the DDGS-supplemented diet could improve growth performance as much as the growth of the control group, and digestibility of dry matter, crude protein, and gross energy were significantly increased by enzyme addition in groups of chicks fed DDGS-supplementation diet. Moreover, the populations of pathogenic bacteria, coliforms, and Bacteroidetes were significantly decreased by enzyme supplementation, which might lead to improved gut mucus-secreting cells and inflammatory cytokines in the jejunum. Collectively, dietary single enzyme and multienzyme complexes could improve gut environments, including intestinal immune responses and gut microbial population, and lead to improvement of growth performance in broiler chickens.

• Journal of Plant Biotechnology
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• 제32권4호
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• pp.233-241
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• 2005

Plants have considerable advantages for the production of antigenic proteins because they provide an inexpensive source of protein and an easy administration of vaccine. Since a publication describing edible plant vaccine of HBsAg in 1992, a number of laboratories around the world have studied the use of plants as the bioreactor to produce antigenic proteins of human or animal pathogens. Over the last ten years, these works have been mainly focused on three major strategies for the production of antigenic proteins in plants: stable genetic transformation of either the nuclear or plastid genome, or transient expression in plants using viral vectors. As many antigenic proteins have been expressed in tobacco, also several laboratories have succeeded to express genes encoding antigenic proteins in other crop plants: potato, tomato, maize, carrot, soybean and spinach. At present many works for the production of edible plant vaccine against bacteria-mediated diseases have mostly performed the studies of enterotoxins and adhesion proteins. Also the development of new-type antigens (pili, flagella, surface protein, other enterotoxin and exotoxin etc.) is required for various targets and more efficacy to immunize against microorganism pathogens. Many works mostly studied in experimental animals had good results, and phase I clinical trial of LTB clearly indicated its immunogenic ability. On the other hand, edible plant vaccines have still problems remained to be solved. In addition to the accumulation of sufficient antigen in plants, human health, environment and agriculture regulation should be proven. Also oral tolerance, the physiological response to food antigens and commensal flora is the induction of a state of specific immunological unresponsiveness, needs to be addressed before plant-derived vaccine becomes a therapeutic option.